Montana State University

Spring 2015

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Mountains and Minds

Written in stone May 12, 2015 by Evelyn Boswell • Published 05/12/15

Seventy-five years after President Franklin D. Roosevelt laid the cornerstone for the Thomas Jefferson Memorial, a microbiologist working at Montana State University visited the scene with a mathematician, architect, conservator, park ranger and scalpel.

Biofilms had made themselves at home on the monument, and Federica Villa, a postgraduate researcher from Italy, had permission from the federal government to investigate. Biofilms, sometimes called slime, are microorganisms whose cells stick together on a surface. In this case, the surface was a national landmark in Washington, D.C.

So, on a stunning day last fall, Villa and her team climbed to the roof of the Thomas Jefferson Memorial and headed toward the discolored areas that indicated biofilms. They spent the day scraping biofilms into test tubes and gathering information about conditions on top of the monument. Then Villa placed the test tubes on dry ice and shipped them overnight to MSU where her colleagues—Betsey Pitts and Breana Pabst—set the biofilms in a freezer at the Center for Biofilm Engineering (CBE).

Storing the biofilms at minus 112 degrees Fahrenheit preserved their genetic material so Villa could analyze them when she returned to Bozeman. Villa is spending two years at the CBE and one year at the University of Milan through a prestigious Marie Curie Fellowship from the European Union.

During her time in the United States, Villa is collecting biofilms four times a year from the Jefferson and Lincoln memorials in Washington, D.C., and the Federal Hall National Memorial in New York City. She is also examining the biofilms in a CBE laboratory on the third floor of MSU’s Engineering and Physical Sciences Building. She can grow the biofilms on small pieces of limestone, for example, and see how they react to acid rain or drought. She can regulate the amount of time they spend in the dark or light.

The goal is to understand what happens on a molecular level when bacteria clump together and form communities on stone, Villa said. She wants to know, too, if biofilms help or harm national treasures made out of stone. People might assume that the effect would be negative, but it’s not necessarily true. The ultimate goal is to help curators clean and preserve art work and monuments without wasting millions of dollars.

Villa is part of the EU-funded ESENCYA project, which was started in 2013 to study the formation of biofilms at the intersection of stone and air, their ability to perceive and respond to environmental cues, and their potential to influence the decay of stone-built treasures, improving restoration and conservation strategies.

“Many of the world’s most precious artworks are made of stone,” according to the project leaders. “Their irreversible deterioration due to biological attack is a worldwide concern. Cyanobacteria colonize outdoor lithic surfaces and develop into biofilms, which, in turn, cause aesthetic, chemical and physical decay. The fragile character of the stone heritage material is further exacerbated by the unpredictable nature of impacts from environmental changes, posing challenges for conservation management.”

Villa—whose work may help lay the groundwork for preventing further decay—said her European supervisor on the project is a worldwide expert on cultural heritage microbiology. Francesca Cappitelli has conducted similar research on the artwork and monuments in Europe.

“What we currently know is that biofilm on stone can give rise to deterioration. However, detecting microorganisms on cultural heritage surfaces does not mean they are damaging the materials,” Cappitelli said. “In addition, the relevance of microbiological aspects to the entire deterioration process should be evaluated carefully for each specific case study.”

Villa’s American supervisor—CBE Director Phil Stewart—said Villa’s project is the first of its kind for the CBE, which already has an international reputation for excellence in biofilm research. He added that he is a strong believer in keeping the CBE flexible so it can respond to new and changing interests.

“Federica is right up there with the most creative and productive postdocs it’s been my pleasure to work with,” said Stewart, who has headed the CBE for almost a decade. “Sometimes people come in with sparks flying, and things happen left and right. She is one of the delightful examples of that.”

Stewart praised Villa’s social skills, as well.

“Federica has just been amazing in her success in networking with people, getting people excited about what she’s working on and collaborating with her,” Stewart added.

Among those scraping biofilms and sharing peanut butter and jelly sandwiches with Villa on the roof of the Jefferson Memorial, for example, was Judy Jacob, senior conservator in the National Park Service’s Northeast Region. Jacob has been an NPS conservator for 28 years and spent a lot of time on roofs, but Villa’s project drew her there again. Even before Villa came to town, Jacob helped her by collecting biofilms on adhesive tape and sending them to Villa so she could examine them under the microscope.

“I have done some studies, but I’m a conservator and not a biologist,” Jacob said. “I have been looking at biofilms, but this is the first time we have had a molecular biologist carrying out a very detailed study of them.”

Another of Villa’s rooftop partners was Isaac Klapper—a former MSU mathematician and longtime CBE collaborator who is now at Temple University in Philadelphia. Already experienced with biofilm fieldwork in Yellowstone National Park, he is on a “very short list of math modelers who are well-known and highly regarded” for their work at the intersection of math and biofilms, Stewart said.

Klapper said he met Jacob in Maine when she was speaking at a conference about “Lichens, Biofilm and Stone.” Seeing the connection to Villa’s work, he introduced the two.

“I don’t know exactly where this project will lead, but it certainly wouldn’t have gotten started without the CBE and the opportunities it cultivates and encourages,” Klapper said.

International reputation

The CBE got its start in 1990 as a National Science Foundation Engineering Research Center. Eleven years later, it graduated from the program and reached the goal of becoming self-sufficient.

Now looking forward to its 25th anniversary in 2015, Stewart said the CBE continues to maintain all three features of an Engineering Research Center: cutting edge interdisciplinary research, innovative education and technology transfer.

“The CBE offers an ideal setting for the interdisciplinary, collaborative research that is the basis for its worldwide reputation in the field of biofilms,” he said. “The CBE has an extraordinary history of outstanding people working together to advance its mission. The combination of creativity, teamwork, excellence and inclusiveness fosters the open environment that leads to shared success.”

Scientists from all over the United States and the world come to the center to conduct research. This year, in addition to Villa, Stewart is supervising postdoctoral researchers from Japan and Brazil. In the past five years, the CBE has hosted 79 visiting researchers from 10 states and 17 foreign countries. They work on a wide variety of projects, ranging from energy production to chronic wounds, from carbon sequestration to Antarctic microbes.

“I like this idea that a small school like MSU, far from urban centers, can draw talent from all over,” Stewart said. “We can be a destination for anybody on the other side of the world to come and work and be creative and collaborate. The pull from the CBE is strong enough that we can draw people internationally.”

Combining science and art

Villa grew up in northern Italy, in a city closer to the chocolates of Switzerland than the Vatican in Rome. Interested in art since she was a child, Villa said microbiology has long intrigued her, as well.

“I was always fascinated by microbiology—something you can’t see with your naked eye, but you know that it’s real and exists,” she said. “I remember when I was a child. I was so curious about small things.”

She went on to earn a master’s degree in environmental science and her Ph.D. in chemistry, biochemistry and ecology of pesticides at the University of Milan, which is a 1 ½-hour train ride away from her hometown of Como. At the University of Milan, she met Cappitelli and realized that cultural heritage microbiology combines her love for art with her love for science.

The two, in fact, worked together on a project involving biofilm staining on the National Museum of the American Indian in Washington, D.C. Cappitelli, Villa and others published a paper about their findings in a 2012 issue of Biofouling: The Journal of Bioadhesion and Biofilm Research.

Villa came to the CBE in 2013. She will leave in August. The landscape and people of Como and Bozeman are enough alike that she feels at home in either place, said Villa, who speaks Italian, English and a little French and loves to cook and bake desserts for her friends. She will then spend the final year of her fellowship at the University of Milan.

Because biofilms also grow on monuments throughout Italy and the rest of Europe, Villa said she will be able to compare their growth on different monuments under different climate conditions.

“For sure, this research may help develop bio-based solutions, something sustainable for the management of cultural heritage,” Villa said. “That would be nice.”

In the meantime, during her remaining time at MSU, Villa will return to Washington, D.C., and New York City to collect more biofilms. A strong believer in collaborations, she said she welcomes the different points of view her team members give her. In addition to her perspective as a microbiologist, they look at biofilms through the eyes of mathematicians, ecologists, engineers and other disciplines. Besides scraping biofilms into test tubes, they gather information about the weather and extreme conditions on top of the monuments.

Biofilms that grow on monuments show a very broad range of tolerance to multiple and fluctuating stresses, Villa said. Temperatures on the monuments change quickly, sometimes going from minus 40 degrees Fahrenheit to 140 degrees Fahrenheit. Water availability fluctuates from long periods of almost total desiccation to being covered with films of water that overlay the biofilm after, for example, a sudden downpour. Also in such hostile environments, the availability of energy sources and nutrients varies from zero to plentiful, like in urban polluted cities.

“These represent extreme conditions for microorganisms,” Villa said. “Although stone inhabitants have to withstand many stresses, they are rarely subject to biological competition, making them exceptional model systems for unraveling symbiotic associations.”

One of Villa’s biofilm photos with Pitts has already made the cover of Microbial Ecology’s “Special Issue on Biofilms.” It was taken with a confocal scanning laser microscope and showed biofilms that colonized a 15th century baptismal font at the Vanderbilt Mansion National Historic Site in Hyde Park, New York. Pitts is a research scientist and microscope facilities manager in the CBE. Anne Camper, a CBE-affiliated member and Regents Professor in MSU’s Department of Civil Engineering, served as guest editor for that July 2014 issue of Microbial Ecology.

While she could have used her fellowship anywhere in the world, the CBE was her first choice, said Villa, who has been a visiting research scholar at Harvard University. The CBE fits her interests perfectly and it’s highly respected, she explained. She also knew what it was like because she spent a few months in the CBE while conducting research for her doctorate.

“The reputation of the CBE is well-known, also in Europe,” Villa said. “It was easy for me to choose my institution.”

Cappitelli agreed, saying the CBE’s reputation in Europe is “a center of excellence, one of the best in the world. Everyone working in the field knows about it.” ■